Aβ1-42 oligomer-induced leakage in an in vitro blood-brain barrier model is associated with up-regulation of RAGE and metalloproteinases, and down-regulation of tight junction scaffold proteins
| Autor: | Chunyan Zhang, Wenbin Wan, Lan Cao, Bill Kalionis, Xiantao Tai, Shijin Xia, Lumei Liu, Yaming Li |
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| Rok vydání: | 2015 |
| Předmět: |
Scaffold protein
Small interfering RNA Blotting Western Receptor for Advanced Glycation End Products In Vitro Techniques Occludin Blood–brain barrier Biochemistry Cell Line Capillary Permeability Mice Cellular and Molecular Neuroscience medicine Animals Humans RNA Small Interfering Receptors Immunologic Receptor Amyloid beta-Peptides Tight Junction Proteins Tight junction Reverse Transcriptase Polymerase Chain Reaction Chemistry Transfection Peptide Fragments Cell biology Endothelial stem cell medicine.anatomical_structure Blood-Brain Barrier Gene Knockdown Techniques Immunology Metalloproteases cardiovascular system |
| Zdroj: | Journal of Neurochemistry. 134:382-393 |
| ISSN: | 0022-3042 |
| Popis: | Accumulating evidence indicates that abnormal deposition of amyloid-β (Aβ) peptide in the brain is responsible for endothelial cell damage and consequently leads to blood–brain barrier (BBB) leakage. However, the mechanisms underlying BBB disruption are not well described. We employed an monolayer BBB model comprising bEnd.3 cell and found that BBB leakage was induced by treatment with Aβ1–42, and the levels of tight junction (TJ) scaffold proteins (ZO-1, Claudin-5, and Occludin) were decreased. Through comparisons of the effects of the different components of Aβ1–42, including monomer (Aβ1–42-Mono), oligomer (Aβ1–42-Oligo), and fibril (Aβ1–42-Fibril), our data confirmed that Aβ1–42-Oligo is likely to be the most important damage factor that results in TJ damage and BBB leakage in Alzheimer's disease. We found that the incubation of bEnd.3 cells with Aβ1–42 significantly up-regulated the level of receptor for advanced glycation end-products (RAGE). Co-incubation of a polyclonal antibody to RAGE and Aβ1–42-Oligo in bEnd.3 cells blocked RAGE suppression of Aβ1–42-Oligo-induced alterations in TJ scaffold proteins and reversed Aβ1–42-Oligo-induced up-regulation of RAGE, matrix metalloproteinase (MMP)-2, and MMP-9. Furthermore, we found that these effects induced by Aβ1–42-Oligo treatment were effectively suppressed by knockdown of RAGE using small interfering RNA (siRNA) transfection. We also found that GM 6001, a broad-spectrum MMP inhibitor, partially reversed the Aβ1–42-Oligo-induced inhibitor effects in bEnd.3 cells. Thus, these results suggested that RAGE played an important role in Aβ-induced BBB leakage and alterations of TJ scaffold proteins, through a mechanism that involved up-regulation of MMP-2 and MMP-9. To reveal the role of RAGE in blood–brain barrier (BBB) disruption in Alzheimer's disease (AD), we employed an monolayer BBB model comprising bEnd.3 cell and found that BBB leakage was induced by treatment with amyloid-β (Aβ), and the levels of tight junction (TJ) scaffold proteins including ZO-1, Claudin-5, and Occludin were decreased. Using receptor for advanced glycation end-products (RAGE) neutralizing polyclonal antibody and siRNA, we confirmed that RAGE played an important role in Aβ-induced BBB leakage and alterations of TJ scaffold proteins and in the up-regulation of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). NF-κB, nuclear factor kappa-light-chain-enhancer of activated B cells. |
| Databáze: | OpenAIRE |
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